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Objective  To assess experimental virulence among sequence type 131 (ST131)  Escherichia coli  bloodstream isolates in relation to virulence genotype and subclone.    Methods  We analysed 48 Spanish ST131 bloodstream isolates (2010) by PCR for ST131 subclone status ( H 30Rx,  H 30 non-Rx, or non- H 30), virulence genes (VGs), and O-type. Then we compared these traits with virulence in a murine sepsis model, as measured by illness severity score (ISS) and rapid lethality (mean ISS ≥ 4).    Results  Of the 48 study isolates, 65% were  H 30Rx, 21%  H 30 non-Rx, and 15% non- H 30; 44% produced ESBLs, 98% were O25b, and 83% qualified as extraintestinal pathogenic  E .  coli  (ExPEC). Of 49 VGs,  ibeA  and  iss  were associated significantly with non- H 30 isolates, and  sat ,  iha  and  malX  with  H 30 isolates. Median VG scores differed by subclone, i.e., 12 ( H 30Rx), 10 ( H 30 non-Rx), and 11 (non- H 30) (p < 0.01). Nearly 80% of isolates represented a described virotype. In mice,  H 30Rx and non- H 30 isolates were more virulent than  H 30 non-Rx isolates (according to ISS [p = 0.03] and rapid lethality [p = 0.03]), as were ExPEC isolates compared with non-ExPEC isolates (median ISS, 4.3 vs. 2.7: p = 0.03). In contrast, most individual VGs, VG scores, VG profiles, and virotypes were not associated with mouse virulence.    Conclusions  ST131 subclone and ExPEC status, but not individual VGs, VG scores or profiles, or virotypes, predicted mouse virulence. Given the lower virulence of non-Rx  H 30 isolates, hypervirulence probably cannot explain the ST131- H 30 clade's epidemic emergence.

plos one

Virulence genes and subclone status as markers of experimental virulence in a murine sepsis model among Escherichia coli sequence type 131 clinical isolates from Spain